Contemporary Research on Bryophytes
By Afroz Alam
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Contemporary Research on Bryophytes - Afroz Alam
In vitro Growth Pattern of Moss Drummondia stricta (Mitt.) Müll. Hal. (Orthotrichaceae) in Different Hormonal Concentrations
Vishwa J. Singh, Dayanidhi Gupta, Vinay Sahu, Ashish K. Asthana*
Bryology Laboratory, CSIR-National Botanical Research Institute, Lucknow-226001, India
Abstract
The present work was undertaken to study the growth pattern of Drummondia stricta (Mitt.) Müll. Hal. in six different culture media with a combination of auxin and cytokinins, using spores as explants in aseptic cultures. In Drummondia, there is a tendency of multicellular spore formation and spore germination is precocious and endosporic. It has been observed that plant growth was the most satisfactory in the ½ Knop’s + 0.1 mg/L IAA + 0.1 mg/L Kinetin followed by ½ Knop’s, 0.1 mg/L IAA + 0.1 mg/L BAP. However, no growth was observed in case of ½ KNOP’s and Hoagland media.
Keywords: Auxin, Cytokinins, Endosporic, Multicellular spores.
* Corresponding author Ashish K. Asthana: Bryology Laboratory, CSIR-National Botanical Research Institute, Lucknow-226001, India; Tel: +91-9415105620; E-mail: drakasthana@rediffmail.com
INTRODUCTION
Genus Drummondia belongs to the family Orthotrichaceae characterized by creeping plants with numerous erect branches. Capsule erect, ovate, exserted with reduced peristome of 16 teeth and spores multicellular, rectangular to polygonal (Plate 1). Vitt [1] described six species and one variety of Drummondia from World. In India 4 species of Drummondia were reported by Chopra [2]. Earlier some research work related to the germination of bryophyte spores, their stages of development, physical factors, nutritional requirements and effects of auxin and antiauxin has been carried out by various workers [3-10]. In bryophytes there are two types of spore germination (1) Endosporic and (2) Exosporic. In endosporic germination, the protonema develop inside the stretched spore wall and the spore protoplast divides within the spore wall forming a multicellular structure while in exosporic germination, swollen protoplast ruptures the spore wall and protonema develops outside. Endosporic germination occurs in some epiphytic and saxicolous species [11]. The aims of the present study were (1) to describe the
spore germination morphology, detailing the phases of in vitro protonema development of D. stricta, and (2) To evaluate the growth of plant in different culture media.
Fig (1))
A-K: Drummondia stricta, A. Plant habit, B-C. Leaves, D. Leaf apical cells, E. Leaf median cells, F. Leaf basal cells, G. Capsule, H. SEM of Spores, I-J. SEM of single spore, K. A portion of spore (enlarged view).
MATERIALS AND METHODS
The plants of D. stricta were obtained from Govind Wildlife Sanctuary, Uttarkashi, Uttarakhand. The specimens have been deposited in the Bryophyte Herbarium of CSIR-National Botanical Research Institute, Lucknow (LWG), India. Specimens examined: India, western Himalaya, Uttarakhand, Uttarkashi, Obra Jeri (alt. ca 2768 m), Epiphytic, 16.10.2016, leg., Dayanidhi Gupta, 306366A (LWG).
The capsules were surface sterilized with 0.5% sodium hypochlorite solution for 2 minutes, and washed repeatedly with sterile double distilled water. The capsules were ruptured and spores were inoculated in different media. The pH of the media was maintained at 5.8 pH before autoclaving. The media was autoclaved at 15 psi for 15 minutes.
The experiment was carried out in a laboratory under controlled temperature (20-23oC) and provided with illumination of 2400-2500 lux as well as alternate light and dark period of 16 h and 8 h, respectively with the help of a combination of fluorescent tubes.
In order to observe the influence of different mineral nutrients and hormone concentration on the development of this species, the following combinations were used. All the hormone concentrations were prepared in ½ Knop's macronutrient.
(A) 0.1 mg/L IBA + 0.2 mg/L BAP + ½ Knop’s
(B) 0.2 mg/L IBA + 0.2 mg/L BAP + ½ Knop’s
(C) 0.1 mg/L 2,4 D + 0.1 mg/L Kinetin + ½ Knop’s
(D) 0.1 mg/L IAA + 0.1 mg/L BAP + ½ Knop’s
(E) Hoagland
(F) ½ Knop’s
RESULTS AND DISCUSSION
Germination Stages and Development
The spores of D. stricta are multicellular with bright green colour with an average size of spores ranging around 80-120 µm long and 60-80 µm wide. In the first week of the analysis, there was development of multicellular protonema enclosed within the spore wall that expanded without rupturing, like a typical endosporic germination. Germination was observed in A, B, C and D media while in the case of Hoagland and ½ Knop’s media only spores were swollen but no further growth occurred. Observation on the development of protonema and sporeling in the second week revealed that in A, B, C and D media massive protonema developed from spores except for Hoagland and ½ Knop’s media. During the fourth week spores and protonema became brown in colour in A, B, C, and D media while in the Hoagland and ½ Knop’s media dead spores were observed (Table 1, Plate 2). The differentiation processes in mosses are clearly dependent on the amount of auxin and cytokinin [12]. In the present experiment cytokinin and auxin accumulation at higher concentrations in the spores inhibited the protonemal growth in A and B media. There are various reports about the effect of auxin and cytokinin on bryophytes, 2, 4 D and 6 Benzyl Amino Purine are usually effective for callus inducement [9, 13, 14]. During the fourth week, leafy buds were observed along the protonema in D media. During the sixth week, leafy bud developed from protonema in C media. Brownish rhizoids also developed from the base of the plants in C and D media. The number of young buds developed more in D media as compared to C media. The plant growth was found best in the case of D media (Table 1, Plate 3). Cvetic et al [15] reported the Auxin IAA favoured budding and the cytokinin 6 Benzyl Amino purine restrained gametophores growth. Both hormones could accelerate protonemal aging in long culture media.
Table 1 Observations on comparative protonemal morphogenesis and growth pattern of D. stricta in different culture media.
Plate (2))
In-vitro growth and multiplication of Drummondia stricta, A-F: Growth of spores in different media after 6 days, A. In medium A, B. In medium B, C. In medium C, D. In medium D, E. In Hoagland medium, F. In ½ Knop’s medium; G-L: Growth of Spores in different media after 15 days, G. In medium A, H. In medium B, I. In medium C, J. In medium D, K. In Hoagland medium, L. In ½ Knop’s medium.
Plate (3))
In-vitro growth and multiplication of D. stricta. Protonema development in different media after 30 days, A-F: A. In medium A, B. In medium B, C. In medium C, D. In medium D, E. In Hoagland medium F. In ½ Knop’s medium; G-J: Protonema and leafy bud development in different media after 45 days, G. In medium A, H. In medium B, I. In medium C, J. In medium D; K-L: Leafy gametophores development after 68 days, K. In medium C, L. In medium D.
CONSENT FOR PUBLICATION
Not applicable.
CONFLICT OF INTEREST
The authors confirm that this chapter contents have no conflict of interest.
ACKNOWLEDGEMENTS
Authors are thankful to the Director, CSIR-National Botanical Research Institute, Lucknow, India for encouraging and providing the facilities.
REFERENCES
Bryophytes and their Associates in South Shetland Islands -Antarctica
Jair Putzke*
University UNIPAMPA, Av. Antonio Trilha, 1847 - São Clemente, São Gabriel - RS, CEP: 97300-000 - Brazil
Abstract
Bryophytes constitute one of the most prominent groups among terrestrial vegetation in South Shetlands Islands of Antarctica. The present study is focused on the bryoflora which exists in association with other life forms in the studied regions. Though, the taxonomy of the Antarctic bryophytes has received much attention in recent years but their associations, especially with lichens and fungi/algae are somewhat neglected by earlier explorers. However, these associations are very crucial for plant succession in the area like Antarctica. A checklist of bryophytes has been provided along with their associated forms with short discussion on the associations to signify the importance of these tiny plants.
Keywords: Antarctica, Associations, Bryophytes, Lichen, Succession.
* Corresponding author Jair Putzke: University UNIPAMPA, Av. Antonio Trilha, 1847 - São Clemente, São Gabriel - RS, CEP: 97300-000 - Brazil; Tel: 51 37177518; E-mail: jrputzkebr@yahoo.com
INTRODUCTION
Antarctica (Plate 1; Fig. 1) was the last continent discovered around 1820 as one of the most fascinating region on the earth with characteristic flora and fauna. At present this remote area of our planet is one of the most drastically changing zones. Climate change is another concern which directly affecting the Antarctic ecosystem in a negative way, affecting the overall quality and quantity of Antarctic species.
Floristic research on Antarctic has received much attention in recent years, especially in context to the influence of climate change and ozone layer depletion on the flora. However, the major research was invariably related to the taxonomy of various plant groups through basic surveys.
The main area of study for bryologists in Antarctica has been the South Shetland Islands (Plate 1; Fig. 2), an archipelago of at least 8 larger islands and dozens of
smaller, where most of the scientific bases are installed and doing research. However, regarding bryophytes, most of the information has been generated in surveys of these islands that resulted in several checklists of Antarctic bryoflora.
Plate 1)
Figure 1. General view of Antarctica; Figure 2. General view of the South Shetland Islands – Elephant Island (Bryorich region).
Ecologically the bryophytes of the Antarctic constitute one of the main components of ecosystem among all the terrestrial organisms present on this continent (Plate 2; Fig. 3). They have important relations with several organisms, being able to be associated with lichens, fungi, algae, higher plants, invertebrates and terrestrial vertebrates. The varied associations denote the importance of these tiny organisms of plant kingdom for the ecology of the studied area.
Plate 2)
Figure 3: Luxuriant growth of terrestrial bryophytes; Figure 4: Sparse growth of bryophytes along with Marchantia; Figure 5: Growth of Cephaloziella in isolated patches; Figure 6: Patch of Pleurocarpous moss Brachythecium; Figures 7 & 8: Patch of acrocarpic